Abstract
To establish a simple and widely accessible technique for rapidly selecting high producing Chinese hamster ovary (CHO) cells engineered to express a monoclonal antibody (mAb), we have exploited the transient display of recombinant protein on their cell surface. In combination with magnetic bead-based methods, we demonstrate the ability to select for cells of high productivity in the absence of any metabolic-based selection method. This technique is sufficient to obtain genetically stable engineered CHO cells via a single step of cell subcloning and yields sought-after stable, high IgG producing clonal cell lines. This technique may also be applied to other types of cells as well as polyclonal Ab cell pools.
Keywords:
Cell line stability; Magnetic cell sorting; Metabolic selection free expression; Recombinant antibody production.
Copyright © 2017 Elsevier B.V. All rights reserved.
MeSH terms
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Animals
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Antibody-Producing Cells / immunology
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Antibody-Producing Cells / metabolism*
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CHO Cells
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Cell Membrane / immunology
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Cell Membrane / metabolism*
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Cloning, Molecular / methods*
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Cricetulus
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Genes, Reporter
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Green Fluorescent Proteins / biosynthesis
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Green Fluorescent Proteins / genetics
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Humans
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Immunoglobulin G / biosynthesis*
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Immunoglobulin G / genetics
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Immunoglobulin G / immunology
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Immunoglobulin Heavy Chains / biosynthesis*
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Immunoglobulin Heavy Chains / genetics
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Immunoglobulin Heavy Chains / immunology
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Immunoglobulin lambda-Chains / biosynthesis*
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Immunoglobulin lambda-Chains / genetics
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Immunoglobulin lambda-Chains / immunology
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Immunomagnetic Separation / methods*
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Phenotype
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / immunology
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Transfection
Substances
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Immunoglobulin G
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Immunoglobulin Heavy Chains
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Immunoglobulin lambda-Chains
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Recombinant Proteins
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Green Fluorescent Proteins